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Methimazole suppresses, and 3-iodothyronamine (T1AM) activates GPCRs, TRs and Na+/K+-ATPase subunit isoform expression in air-breathing fish hearts 甲巯咪唑抑制呼吸鱼心脏gpcr、TRs和Na+/K+- atp酶亚基异构体的表达,而3-碘thyronamine (T1AM)则激活gpcr、TRs和Na+/K+- atp酶亚基异构体的表达
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-22 DOI: 10.1016/j.ygcen.2025.114802
P.U. Ushas , M.C. Subhash Peter
3-iodothyronamine (T1AM), an endogenous aminergic derivative of thyroid hormone (TH), exhibits cardioprotective effects in mammals. However, it is unclear whether T1AM has similar effects in fish hearts to counteract hypothyroidism. We hypothesise that T1AM may regulate the expression patterns of GPCR-associated neuroendocrine receptors, thyroid hormone receptors (TRs), and Na+, K+-ATPase (NKA) subunit isoform genes, and may exert protection against hypothyroidism. The effects of T1AM on ion transporter activities in fish hearts under euthyroid conditions were first examined in vitro and in vivo to ascertain its direct impact on euthyroid fish hearts. Subsequently, the effects of methimazole (MMI), an antithyroid drug, and T1AM replacement in hypothyroid hearts of the air-breathing fish (Anabas testudineus) were studied. Dose-dependent effects of T1AM on ion-dependent ATPase activities were observed both in vitro and in vivo in euthyroid hearts. While MMI treatment increased NKA activity, T1AM replacement decreased its activity in hypothyroid hearts. The downregulation of NKA subunit isoform expressions (nkaα1a, nkaα1b, nkaα1c, atp1b1, atp1b2) by MMI was reversed when T1AM was added in hypothyroid hearts. Similarly, MMI suppressed and T1AM activated the transcript of trace amine-associated receptors taar1 and taar8c, as well as TR isoforms (thra, thrab, thrb) in the hypothyroid heart. MMI activated adra2a and suppressed adrb2 expression, while T1AM reversed these effects; however, both MMI and T1AM downregulated drd2 expression. These data indicate a novel targeted action of T1AM on cardiac GPCRs, TR function, and NKA-mediated ion osmotic homeostasis in hypothyroid fish, suggesting potential cardioprotective effects of T1AM against hypothyroidism.
3-碘甲状腺胺(T1AM)是甲状腺激素(TH)的内源性胺类衍生物,在哺乳动物中具有心脏保护作用。然而,尚不清楚T1AM是否在鱼类心脏中有类似的作用来对抗甲状腺功能减退。我们假设T1AM可能调节gpcr相关的神经内分泌受体、甲状腺激素受体(TRs)和Na+, K+- atp酶(NKA)亚基异构体基因的表达模式,并可能对甲状腺功能减退起到保护作用。首先在体内和体外研究了T1AM对正常甲状腺鱼心脏离子转运体活性的影响,以确定其对正常甲状腺鱼心脏的直接影响。随后,研究了抗甲状腺药物甲巯咪唑(MMI)和T1AM替代对呼吸鱼(Anabas testudineus)甲状腺功能减退心脏的影响。在体内和体外观察了T1AM对正常甲状腺心脏离子依赖性atp酶活性的剂量依赖性影响。虽然MMI治疗增加了NKA活性,但T1AM替代降低了甲状腺功能减退心脏的NKA活性。在甲状腺功能减退的心脏中加入T1AM后,MMI下调NKA亚基亚型(NKA α1a、NKA α1b、NKA α1c、atp1b1、atp1b2)的表达逆转。同样,MMI抑制和T1AM激活了甲状腺功能减退心脏中微量胺相关受体taar1和taar8c以及TR异构体(thra, thrab, thrb)的转录。MMI激活adra2a并抑制adrab2的表达,而T1AM逆转了这些作用;然而,MMI和T1AM都下调了drd2的表达。这些数据表明,T1AM对甲状腺功能减退鱼的心脏gpcr、TR功能和nka介导的离子渗透稳态具有新的靶向作用,表明T1AM对甲状腺功能减退具有潜在的心脏保护作用。
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引用次数: 0
Effects of mineralocorticoids on α-Na+/K+ ATPase expression and parasite morphology in T. crassiceps WFU cysticerci 矿化皮质激素对柞蚕WFU囊虫α-Na+/K+ atp酶表达及形态的影响
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-22 DOI: 10.1016/j.ygcen.2025.114803
Jesus Angel Aguirre-Pineda , Ricardo Arturo Valdéz Perez , Alfredo Pérez-Guiot , Raúl J. Bobes , Marta Catalina Romano
Mineralocorticoids are corticosteroid hormones that, in vertebrates, regulate extracellular fluid volume and salt balance by modulating the expression and activity of Na+/K+ ATPase, the enzyme that generates and maintains Na+ and K+ gradients across plasma membranes. However, the function of mineralocorticoids in invertebrates has not been well studied. Cysticerci (larvae) of the parasite Taenia crassiceps WFU synthesize the corticosteroids 11-deoxycorticosterone (DOC) and corticosterone in vitro. Here, we searched the genome and transcriptome of T. crassiceps for the presence of α-Na+/K+ ATPase sequences. We also studied the effects of corticosteroids on α-Na+/K+ ATPase expression and on the morphology of T. crassiceps WFU cysticerci in vitro. Cysticerci were obtained from the peritoneal cavity of previously infected female mice and cultured in the presence of corticosterone, DOC, spironolactone (a mineralocorticoid receptor antagonist) or metyrapone (a corticosteroid synthesis inhibitor) at different concentrations and for different periods of time. ATPase expression was evaluated by Western blotting. Genome and transcriptome searches revealed two highly conserved α-Na+/K+ ATPase sequences in T. crassiceps WFU. DOC was found to increase α-Na+/K+ ATPase expression in a concentration-dependent manner. The incubation of parasites with spironolactone, but not with metyrapone, significantly decreased the expression of the enzyme. In addition, DOC increased parasite size, whereas spironolactone and metyrapone altered parasite shape and size, respectively. These results suggest a role for mineralocorticoids in salt–water regulation of T. crassiceps WFU. A deep understanding of these mechanisms could contribute to improving therapeutic strategies for cysticercosis.
矿化皮质激素是一种皮质类固醇激素,在脊椎动物中,通过调节Na+/K+ atp酶的表达和活性来调节细胞外液的体积和盐的平衡。Na+/K+ atp酶是一种产生和维持Na+和K+在质膜上梯度的酶。然而,矿物皮质激素在无脊椎动物中的作用尚未得到很好的研究。绦虫带绦虫囊虫(幼虫)体外合成皮质甾体11-脱氧皮质酮(DOC)和皮质酮。在此,我们检索了T. crassiceps的基因组和转录组,寻找α-Na+/K+ atp酶序列的存在。我们还研究了糖皮质激素对体外培养的天蚕囊尾蚴α-Na+/K+ atp酶表达和形态的影响。从先前感染的雌性小鼠的腹膜腔中获得囊虫,并在皮质酮、DOC、螺内酯(一种矿物皮质激素受体拮抗剂)或美替拉酮(一种皮质类固醇合成抑制剂)存在下以不同浓度和不同时间进行培养。Western blotting检测ATPase表达。通过基因组和转录组搜索,我们发现了两条高度保守的α-Na+/K+ atp酶序列。DOC可使α-Na+/K+ atp酶的表达呈浓度依赖性增加。与螺内酯孵育的寄生虫,而不是与metyrapone显著降低酶的表达。此外,DOC增加了寄生虫的大小,而螺内酯和metyrapone分别改变了寄生虫的形状和大小。这些结果提示矿物皮质激素在水盐调控中起一定作用。深入了解这些机制有助于改进囊虫病的治疗策略。
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引用次数: 0
Multi-year profiles of T3 are positively correlated with corticosterone in male bowhead whale baleen 雄性弓头鲸须中T3的多年谱与皮质酮呈正相关。
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-18 DOI: 10.1016/j.ygcen.2025.114800
Justine M. Hudson , Kathleen Hunt , Marianne Marcoux , C.Loren Buck , Cory J.D. Matthews
Thyroid hormones play an important role in the regulation of growth, development, metabolism, thermoregulation, and migration. Very little information exists on patterns of thyroid hormone concentrations in healthy mysticete whales, as many studies have focused on ill, entangled, or stranded whales, making it difficult to interpret thyroid hormone trends. In this study, we used a unique sample-set of bowhead whale baleen plates to explore the long-term interrelationships between triiodothyronine (T3), the most biologically active thyroid hormone, corticosterone, testosterone, and nitrogen isotope ratios (δ15N) (proxies for stress, reproduction, and diet, respectively) to investigate the role T3 may play in the physiology of healthy cetaceans. Baleen plates were collected between 1998 and 2011 from eight subsistence-harvested male bowhead whales across the Eastern Canadian Arctic. Each baleen plate generated 88–158 serial samples, representing ∼11–22 years of life for each individual whale. T3 concentrations ranged from 0.61 to 21.62 ng/g and varied seasonally in just two whales. Most whales showed no correlation between T3 and seasonal fluctuations in testosterone or δ15N, suggesting that variation in T3 is not driven by seasonal shifts in reproductive cycles, consumer trophic level, or migration. However, a strong positive correlation between T3 and corticosterone was observed in every whale, which we hypothesized was due to non-seasonal factors that simultaneously increase metabolic rate and physiological stress. The positive correlation between T3 with corticosterone suggests that in mysticete whales, some stressors may require increased energetic output.
甲状腺激素在调节生长、发育、代谢、体温调节和迁移等方面发挥重要作用。关于健康神秘鲸体内甲状腺激素浓度模式的信息很少,因为许多研究都集中在生病、被纠缠或搁浅的鲸鱼身上,因此很难解释甲状腺激素的趋势。在这项研究中,我们利用一组独特的弓头鲸须骨板样本,探索了三碘甲状腺原氨酸(T3),最具生物活性的甲状腺激素,皮质酮,睾酮和氮同位素比率(δ15N)(分别代表压力,繁殖和饮食)之间的长期相互关系,以研究T3在健康鲸类生理中的作用。1998年至2011年间,研究人员从加拿大东部北极地区的8头雄性弓头鲸身上收集了鲸须片。每个鲸须板产生88-158个连续样本,代表每只鲸鱼的寿命~ 11-22 年。T3浓度范围为0.61-21.62 ng/g,仅两只鲸鱼的浓度随季节变化。大多数鲸鱼的T3与睾酮或δ15N的季节性波动之间没有相关性,这表明T3的变化不是由生殖周期、消费者营养水平或迁徙的季节性变化驱动的。然而,在每只鲸鱼中,T3和皮质酮之间都观察到很强的正相关,我们假设这是由于非季节性因素同时增加了代谢率和生理应激。T3与皮质酮之间的正相关表明,在神秘鲸中,一些压力源可能需要增加能量输出。
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引用次数: 0
Catecolaminergic systems and hypothalamic-pituitary-thyroid (HPT) axis: adrenaline, noradrenaline and dopamine treatments in vivo in lizard Podarcis siculus 儿茶酚胺能系统和下丘脑-垂体-甲状腺(HPT)轴:肾上腺素、去甲肾上腺素和多巴胺在蜥蜴体内的作用。
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-16 DOI: 10.1016/j.ygcen.2025.114801
Rosaria Sciarrillo , Alessandra Falzarano , Vito Gallicchio , Assunta Lallo , Francesca Carrella , Aldo Mileo , Anna Capaldo , Maria De Falco
The catecolaminergic system, which uses dopamine, noradrenaline and adrenaline as neurotransmitters, originates in the brainstem nuclei and spreads widely throughout the central nervous system (CNS). Although there are data indicating that interactions between the catecolaminergic systems of the nervous system and hypothalamic-pituitary-thyroid axis do indeed take place and may be of physiological and clinical relevance, such interactions are far from being clarified. In this paper, we studied the hypothalamic-pituitary-thyroid axis responses in lizards treated with adrenaline, noradrenaline and dopamine respectively. Adrenaline raised plasma TRH and TSH levels and induced a stimulatory effect on the thyroid gland activity with an increase of T3 and T4 levels. On the contrary, noradrenaline treatment induced a reduction in TRH secretion, accompanied by a normal circulating TSH level and increased plasma T4 levels but with a reduction of circulating T3 levels. Dopamine stimulated TRH secretion centrally but induced TSH deficiency followed by reduction of circulating T3 and T4 levels. Furthermore, the effects of the catecolaminergic system on the TRH-TSH-thyroid hormone axis has been also manifested through mechanisms in peripheral organs such as the liver. The dynamic interplay between TRH and TSH and the feedback effects on circulating thyroid hormones after treatment with adrenaline, noradrenaline and dopamine is a very sophisticated and complicated mechanism that responds via afferent inputs from neurons originating in the paraventricular nucleus and can influence the secretion of TRH and TSH-secreting hypophysiotropic neurons with consequent feedback on thyroid hormones.
儿茶酚胺能系统以多巴胺、去甲肾上腺素和肾上腺素为神经递质,起源于脑干核,并在整个中枢神经系统(CNS)中广泛传播。虽然有数据表明神经系统儿茶酚胺能系统与下丘脑-垂体-甲状腺轴之间的相互作用确实发生,并且可能具有生理和临床相关性,但这种相互作用远未被阐明。本文分别研究了肾上腺素、去甲肾上腺素和多巴胺对蜥蜴下丘脑-垂体-甲状腺轴的影响。肾上腺素升高血浆TRH和TSH水平,并通过增加T3和T4水平诱导甲状腺活性的刺激作用。相反,去甲肾上腺素治疗导致TRH分泌减少,同时循环TSH水平正常,血浆T4水平升高,但循环T3水平降低。多巴胺刺激TRH中枢分泌,但诱导TSH缺乏,随后循环T3和T4水平降低。此外,儿茶酚胺能系统对trh - tsh -甲状腺激素轴的影响也通过肝脏等外周器官的机制表现出来。肾上腺素、去甲肾上腺素和多巴胺治疗后,TRH和TSH之间的动态相互作用以及对循环甲状腺激素的反馈作用是一个非常复杂的机制,它通过源自室旁核的神经元的传入输入来响应,并影响TRH和分泌TSH的垂体性神经元的分泌,从而对甲状腺激素产生反馈。
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引用次数: 0
Neurosecretory protein GL stimulates feeding behavior and fat accumulation in Japanese quails (Coturnix japonica) 神经分泌蛋白GL刺激日本鹌鹑摄食行为和脂肪积累
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-11 DOI: 10.1016/j.ygcen.2025.114798
Masaki Kato, Eiko Iwakoshi-Ukena, Megumi Furumitsu, Yuki Narimatsu, Kazuyoshi Ukena
Neurosecretory protein GL (NPGL) is a novel hypothalamic neuropeptide that promotes fat accumulation in rats, mice, and neonatal chicks. However, its role in Japanese quail (Coturnix japonica) remains unclear. In this study, we investigated the effects of chronic intracerebroventricular infusion of NPGL on body mass, food intake, and fat deposition in five-week-old male Japanese quails. A 13-day infusion of NPGL significantly increased body mass, food intake, and the masses of subcutaneous fat, abdominal fat, and liver. In contrast, water intake and the masses of the pancreas, testes, heart, and muscle remained unchanged. Blood concentrations of triglyceride, glucose, and non-esterified fatty acid were unaffected. Real-time qPCR analysis revealed a significant upregulation of NPGM, a paralogous gene of NPGL, in the hypothalamus. Additionally, the expression of stearoyl-CoA desaturase 1 (SCD1), a key enzyme involved in lipogenesis, showed an increasing trend in the liver. Although the fatty acid ratio used as the SCD1 activity index (palmitoleate/palmitate) did not change, the de novo lipogenesis index (palmitate/linoleate) tended to increase in both the liver and abdominal fat. These results suggest that NPGL promotes fat accumulation in Japanese quails, indicating its potential involvement in avian adiposity.
神经分泌蛋白GL (NPGL)是一种新的下丘脑神经肽,可促进大鼠、小鼠和新生雏鸡的脂肪积累。然而,它在日本鹌鹑(Coturnix japonica)中的作用尚不清楚。在这项研究中,我们研究了慢性脑室内灌注NPGL对5周龄雄性日本鹌鹑体重、食物摄入和脂肪沉积的影响。13天的NPGL输注显著增加了体重、食物摄入量以及皮下脂肪、腹部脂肪和肝脏的体积。相比之下,水的摄入量和胰腺、睾丸、心脏和肌肉的质量保持不变。血液中甘油三酯、葡萄糖和非酯化脂肪酸的浓度未受影响。Real-time qPCR分析显示,NPGM是NPGL的一个副同源基因,在下丘脑中显著上调。此外,肝脏中参与脂肪生成的关键酶硬脂酰辅酶a去饱和酶1 (SCD1)的表达呈上升趋势。虽然用作SCD1活性指数的脂肪酸比率(棕榈油酸酯/棕榈酸酯)没有变化,但肝脏和腹部脂肪的新生脂肪生成指数(棕榈酸酯/亚油酸酯)都有增加的趋势。这些结果表明,NPGL促进日本鹌鹑的脂肪积累,表明其可能参与鸟类肥胖。
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引用次数: 0
Regulation of transforming growth factor beta 1 in the ovary of ovoviviparous black rockfish (Sebastes schlegelii) 卵胎生黑岩鱼卵巢转化生长因子β 1的调控
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-10 DOI: 10.1016/j.ygcen.2025.114799
Xiao Jing, Likang Lyu, Chenpeng Zuo, Jianshuang Li, Xiaojie Wang, Jing Yang, Tianyu Jiang, Yun Li, Haishen Wen, Xin Qi
Transforming growth factor beta 1 (TGF-β1), a multifunctional secreted polypeptide cytokine, has been reported to play crucial roles in pregnancy process of mammals, including immune tolerance and embryonic development. However, less is known in Pisces. Aimed to figure out the molecular mechanism underlying TGF-β1 functions, black rockfish (Sebastes schlegelii), an ovoviviparity teleost, which process the sperm storage and gestation periods, was employed as the research model. In the present study, we found that tgfb1 among four tgfb isoforms was highly expressed and localized in ovary. The recombinant TGF-β1 (rTGF-β1) was mainly involved in immune response, signal transduction, angiogenesis and cell death by analyzing transcriptome of ovarian cells. The current results reveal the pivotal role of TGF-β1 in black rockfish ovary and provide novel insights into cytokines in the pregnancy of ovoviviparity teleost.
转化生长因子β1 (TGF-β1)是一种多功能分泌型多肽细胞因子,据报道在哺乳动物妊娠过程中,包括免疫耐受和胚胎发育中起着至关重要的作用。然而,对双鱼座的了解却很少。为了弄清TGF-β1功能的分子机制,我们以卵生硬骨鱼黑岩鱼(sebases schlegelii)作为研究模型,黑岩鱼是一种处理精子储存和妊娠期的硬骨鱼。在本研究中,我们发现tgfb四种亚型中tgfb1在卵巢中高度表达和定位。重组TGF-β1 (rTGF-β1)主要参与卵巢细胞的免疫应答、信号转导、血管生成和细胞死亡。本研究结果揭示了TGF-β1在黑岩鱼卵巢中的关键作用,并为细胞因子在卵生硬骨鱼妊娠中的作用提供了新的见解。
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引用次数: 0
Scale cortisol reflects the acute but not the chronic stress status in green sunfish (Lepomis cyanellus) 鳞皮质醇反映了绿翻车鱼(Lepomis cyanellus)的急性而非慢性应激状态。
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-08-09 DOI: 10.1016/j.ygcen.2025.114797
Abigayle M. McKinley, Carol Best, Nicholas J. Bernier
Scale cortisol has the potential to be a useful biomarker of chronic stress in fish, but the dynamics of scale cortisol accumulation and release are not well understood. In this study, we assessed the dynamics of plasma and scale cortisol accumulation and clearance in green sunfish (Lepomis cyanellus) in response to acute and chronic stressors. While sunfish exposed to 21 days of unpredictable chronic stress (UCS) mounted an attenuated acute plasma cortisol response to the daily stressor on day 21 and had reduced liver and body conditions, their scale cortisol content was either unchanged or reduced. In contrast, exposure to an acute stressor increased scale cortisol content 30 min post-stressor in naïve fish but had no effect when preceded by the 21-day UCS protocol. We also examined the effects of an acute stressor on scale cortisol content in goldfish (Carassius auratus) to identify potential species differences in scale cortisol dynamics. Interestingly, although an acute stressor increased scale cortisol content 30 min post-stressor and declined 24 h later in both species, only sunfish scale cortisol levels returned to baseline by 24 h post-stressor. Together, our results indicate that scales in green sunfish can accumulate and clear cortisol rapidly following an acute stressor and do not reflect long-term accumulation of cortisol from the plasma during chronic stress. As such, we conclude that scale cortisol content is not a biomarker of chronic stress in green sunfish.
鳞片皮质醇有可能成为鱼类慢性应激的有用生物标志物,但鳞片皮质醇积累和释放的动力学尚不清楚。在这项研究中,我们评估了绿色太阳鱼(Lepomis cyanellus)在急性和慢性应激源下血浆和鳞片皮质醇积累和清除的动态。虽然太阳鱼暴露在21 天的不可预测的慢性应激(UCS)中,第21天对日常应激源的急性血浆皮质醇反应减弱,肝脏和身体状况下降,但它们的鳞片皮质醇含量不变或降低。相比之下,暴露于急性应激源增加了naïve鱼的鳞片皮质醇含量30 min后应激源,但在21天的UCS方案之前没有影响。我们还研究了急性应激源对金鱼(Carassius auratus)体表皮质醇含量的影响,以确定体表皮质醇动态的潜在物种差异。有趣的是,尽管急性应激源在应激后30 min增加了两种物种的皮质醇水平,并在应激后24 h下降,但只有太阳鱼的皮质醇水平在应激后24 h恢复到基线水平。总之,我们的研究结果表明,绿色太阳鱼的鳞片可以在急性应激源后迅速积累和清除皮质醇,而不反映慢性应激期间血浆中皮质醇的长期积累。因此,我们得出结论,尺度皮质醇含量不是绿色太阳鱼慢性应激的生物标志物。
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引用次数: 0
Disorders of glucose metabolism and impaired energy metabolism in pfkma and pfkmb knockout zebrafish pfkma和pfkmb敲除对斑马鱼糖代谢紊乱和能量代谢的影响。
IF 1.7 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-07-23 DOI: 10.1016/j.ygcen.2025.114787
Na Liu , Mengjie Li , Tianci Liu , Jitu Wang , Bo Zhang , Man Zhang

Background

In our previous study, Phosphofructokinase, muscle (PFKM), a key enzyme in glucose metabolism, was found to be decreased in the urinary exosomes of diabetic patients. The role of PFKM in glucose and energy metabolism is not fully understood. The study explores PFKM’s implications in these metabolic pathways through the lens of a zebrafish model.

Methods

We analyzed pfkma and pfkmb expressions in early zebrafish development using Daniocell and ZESTA databases and performed whole-mount in situ hybridization on embryos up to 4 days post-fertilization (dpf). The CRISPR/Cas9 system was utilized to generate pfkma and pfkmb knockout zebrafish, and the expression of various genes, including β-actin, pfkma, pfkmb, peo1, mfn1, drp1, nd2, cytb, cox3, and atp6, was measured by qRT-PCR in the embryos.

Results

Knockout mutants displayed elevated glucose levels across whole embryos. Additionally, at 5 dpf, pfkma and pfkmb double mutants showed pancreatic islet proliferation, ATP depletion, downregulation of the expression of the mitochondrial-related gene peo1, and impaired locomotor activity.

Conclusions

PFKM plays an important role in stabilizing glucose metabolism and has a certain impact on mitochondrial energy metabolism pathways in zebrafish model.
背景:在我们之前的研究中,发现糖尿病患者尿外泌体中葡萄糖代谢的关键酶磷酸果糖激酶(PFKM)减少。PFKM在葡萄糖和能量代谢中的作用尚不完全清楚。该研究通过斑马鱼模型探讨了PFKM在这些代谢途径中的影响。方法:利用Daniocell和ZESTA数据库分析pfkma和pfkmb在斑马鱼早期发育过程中的表达,并对4天大的胚胎进行全载原位杂交。利用CRISPR/Cas9系统构建pfkma和pfkmb敲除斑马鱼,qRT-PCR检测胚胎中β-actin、pfkma、pfkmb、peo1、mfn1、drp1、nd2、cytb、cox3和atp6等多种基因的表达。结果:敲除突变体在整个胚胎中显示葡萄糖水平升高。此外,在受精后5 天(dpf), pfkma和pfkmb双突变体表现出胰岛增生,ATP水平下降,线粒体相关基因peo1表达减少,运动活性降低。结论:PFKM在稳定斑马鱼模型葡萄糖代谢中具有重要作用,并对线粒体能量代谢途径有一定影响。
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引用次数: 0
Ontogeny of gonadotropin-inhibitory hormone cells and fibers in the brain and pituitary gland of the pejerrey fish, Odontesthes bonariensis 鱼脑和脑垂体中促性腺激素抑制激素细胞和纤维的个体发育
IF 2.1 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-07-18 DOI: 10.1016/j.ygcen.2025.114788
Ana M. Pahí-Rosero , Gabriela C. López , María R. Pérez , Alejandro S. Mechaly , Takayoshi Ubuka , José A. Muñoz-Cueto , Paula G. Vissio , Gustavo M. Somoza
The gonadotropin-inhibitory hormone is a neuropeptide belonging to the RF-amide family of peptides, first characterized in birds. This peptide can inhibit the synthesis and release of gonadotropins in both avians and mammals. Although gonadotropin-inhibitory hormone (Gnih) variants have also been detected in different fish species, knowledge of their physiological action is still inconclusive and controversial. In addition, there is very little information on the neuronal development of Gnih cells in early stages. In this context, the objective of the present study is to characterize Gnih-containing neurons and fibers in the brain and pituitary gland of adult fish, and during early development of the pejerrey (Odontesthes bonariensis), and the possible relationship with the sex differentiation process.
The Gnih neurons and fibers were determined by immunohistochemistry by using different antisera in the adult pejerrey brain. Gnih-immunoreactive (Gnih-ir) neurons were observed in the olfactory bulbs, the terminal nerve ganglion, the lateral nuclei of the ventral telencephalon, the posterior periventricular nucleus, the dorsal tegmentum, and some isolated neuronal bodies were detected in the secondary gustatory nucleus. Very few Gnih-ir fibers were detected innervating the pituitary gland.
The Gnih neuronal and fiber distribution was also studied from hatching to week 10, covering the sex differentiation period until the gonadal sex was observed. Gnih-immunoreactive neuronal bodies were identified from hatching in the terminal nerve ganglion and the diencephalic posterior periventricular nucleus. Gnih-ir fibers were observed in many regions, including the retina, and a profuse innervation was observed in the pituitary. From the first week post-hatching, Gnih-ir neuronal bodies were identified in the dorsal mesencephalic tegmentum and the lateral nucleus of the ventral telencephalon. In addition, Gnih-ir cells were detected in the pituitary. These Gnih-ir cells were consistently detected from 3 to 7 weeks after hatching, coinciding with onset of gonadal differentiation. At week 10 and in the adult, only a few Gnih-ir fibers were observed in the pituitary. Although the precise function of these pituitary Gnih-ir cells is unknown, the relationship between the appearance of these cells and the process of sex differentiation suggests that Gnih may influence this process.
促性腺激素抑制激素是一种神经肽,属于rf -酰胺家族的肽,首次在鸟类中被发现。这种肽可以抑制鸟类和哺乳动物促性腺激素的合成和释放。虽然在不同的鱼类中也检测到促性腺激素抑制激素(Gnih)变异,但其生理作用的知识仍然是不确定和有争议的。此外,关于Gnih细胞早期神经元发育的信息很少。在此背景下,本研究的目的是表征成年鱼和bonariensis (Odontesthes bonariensis)早期发育过程中大脑和垂体中含有gnh的神经元和纤维,以及它们与性别分化过程的可能关系。采用免疫组化方法,用不同抗血清对成年大鼠脑Gnih神经元和纤维进行了检测。在嗅球、末梢神经节、脑端腹侧外侧核、脑室后周核、背被中可见Gnih-ir免疫反应神经元,在次级味觉核中可见部分孤立的神经元体。脑下垂体的gnh -ir神经纤维分布较少。从孵化到第10周,研究Gnih神经元和纤维的分布,包括性别分化期,直到观察性腺性别。从末梢神经节和间脑后室周核中孵化出gnh免疫反应性神经元体。在包括视网膜在内的许多区域均可见gnh -ir纤维,在垂体中可见丰富的神经支配。从孵化后第一周开始,在中脑背被和端脑腹侧外侧核中发现了gnh -ir神经元体。此外,垂体中检测到Gnih-ir细胞。这些gnh -ir细胞在孵化后3至7周持续检测到,与性腺分化的开始一致。在第10周和成人中,仅在垂体中观察到少量gnh -ir纤维。虽然这些垂体Gnih-ir细胞的确切功能尚不清楚,但这些细胞的出现与性别分化过程之间的关系表明,Gnih可能影响这一过程。
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引用次数: 0
In vitro effects of polycyclic aromatic hydrocarbons on sex steroid production by Arabian sea bream Acanthopagrus arabicus ovarian cells 多环芳烃对阿拉伯海棘鱼卵巢细胞产生性类固醇的体外影响
IF 2.1 3区 医学 Q3 ENDOCRINOLOGY & METABOLISM Pub Date : 2025-07-05 DOI: 10.1016/j.ygcen.2025.114778
Fereshteh Aysham , Negin Salamat , Alireza Safahieh , Arash Larki , Asma Mohammadi
Polycyclic aromatic hydrocarbons (PAHs), considered endocrine disruptors, affect the animals reproduction by interfering with the production of sex steroids. In the present study, the in vitro effects of selected PAHs (naphthalene, phenanthrene and BaP) on steroid production of cultivated ovarian cells from Arabian sea bream (Acanthopagrus arabicus) was assessed. Ovarian cells were cultured in Leibovitz L-15 (L-15) medium containing naphthalene, phenanthrene and benzo(a)pyrene with or without androstenedione (AD) as precursors and17b-estradiol (E2), progesterone (P4) and testosterone (T) were then measured. The effects of selected PAHs on the steroidogenic enzyme systems cytochrome P450 17, 20-lyase (P450-17,20l) and cytochrome P450 aromatase (P450-arom) were also compared with the ketoconazole (KCZ) action as inhibitor of cytochrome P450 steroidogenic enzymes (cytochrome P450 17, 20-lyase (P450-17,20l)) and cytochrome P450 aromatase (P450-arom). Adding exogenous androstenedione as a steroid precursor significantly increased the production of all three steroid hormones by cultured ovarian cells. On the other hand, the addition of ketoconazole significantly reduced the production of E2 and P from ovarian cells, while it had no effect on the production of T. This result showed that ketoconazole only affects the cytochrome P450 steroidogenic enzymes and has no effect on the 17-beta-hydroxysteroid dehydrogenase (17-βHSD) enzyme that convert androstenedione to testosterone. On the other hand, a significant decrease in the production of all three steroids (17β-estradiol, progesterone, and testosterone) from ovarian cells exposed to the studied pollutants (naphthalene, phenanthrene, and benzo(a)pyrene) even in the presence of androstenedione indicated that PAHs inhibit all three steroidogenic enzyme systems including P450-17,20l, P450-arom, and 17b-HSD. In conclusion, PAHs are potent inhibitors of the steroidogenic enzyme system including P450-17,20l, P450-arom, and 17b-HSD, and therefore, they can disturb the reproduction of fish living in contaminated areas due to impairment of steroid biosynthesis.
多环芳烃(PAHs)被认为是内分泌干扰物,它通过干扰性激素的产生来影响动物的生殖。本研究研究了选定的多环芳烃(萘、菲和BaP)对阿拉伯海鲷(Acanthopagrus arabicus)卵巢细胞类固醇生成的体外影响。卵巢细胞在含萘、菲和苯并芘(a)芘的Leibovitz L-15 (L-15)培养基中培养,分别以或不以雄烯二酮(AD)为前体,并测定17b-雌二醇(E2)、孕酮(P4)和睾酮(T)。比较了所选多环芳烃对细胞色素P450 17,20 -裂解酶(P450-17,20l)和细胞色素P450芳香化酶(P450-arom)的影响,并比较了酮康唑(KCZ)作为细胞色素P450甾体化酶(细胞色素P450 17,20 -裂解酶(P450-17,20l))和细胞色素P450芳香化酶(P450-arom)抑制剂的作用。添加外源性雄烯二酮作为类固醇前体,可显著增加培养卵巢细胞三种类固醇激素的产生。另一方面,酮康唑的加入显著降低卵巢细胞E2和P的产生,而对t的产生没有影响。这表明酮康唑只影响细胞色素P450甾体生成酶,而对将雄烯二酮转化为睾酮的17-β -羟基类固醇脱氢酶(17-βHSD)没有影响。另一方面,即使在雄烯二酮存在的情况下,暴露于所研究的污染物(萘、菲和苯并(a)芘)的卵巢细胞中所有三种类固醇(17β-雌二醇、孕酮和睾酮)的产生也显著减少,这表明多环芳烃抑制了所有三种类固醇生成酶系统,包括P450-17、20l、p450 -芳烃和17b-HSD。综上所示,多环芳烃是P450-17、20l、P450-arom和17b-HSD等甾体生成酶系统的有效抑制剂,可以通过抑制甾体合成来干扰污染区鱼类的繁殖。
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General and comparative endocrinology
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